1. Technical Field
The present invention relates to a probe for detecting polymorphism, a method of detecting polymorphism, a method of evaluating a drug efficacy or tolerance, a disease prediction method, and a reagent kit for detecting polymorphism.
2. Related Art
So far, Urate Transporter (URAT1/SLC22A12), which is a uric acid reabsorption transporter, and Glucose Transporter 9 (GLIT9/SLC22A12) have been known as therapeutic target molecules for hyperuricemia.
In recent years, it has been proved that the ABCG2 (ATP-binding cassette sub-family G member 2) gene, which is also referred to as BCRP (breast cancer resistance protein), codes for a high volume urination transporter.
It has also been suggested that the presence of polymorphism in the ABCG2 gene is linked to an increased risk of developing gout, and it has been proved that the ABCG2 gene is a causative gene of gout (see, for example, JIKKEN IGAKU (Experimental Medicine), 2010, Vol. 28, No. 8, pp. 1285-1289).
Further, it has been suggested that the presence of polymorphism in the ABCG2 gene is relevant to abnormal protein expression in a placenta (see, for example, Drug. Metab. Dispos., 2005, Vol. 33, No. 1, pp. 94-101), and a method, in which polymorphism in the ABCG2 gene may be measured in a short time, at a low cost and easily, is demanded.
As a method of measuring polymorphism of a gene, a PCR-RFLP method is known. In this method, PCR is carried out using primers that have been designed so as to amplify a region containing bases that are desired to be measured; the products obtained by the amplification are subjected to cleaving with a restriction enzyme, that is selected so that the presence or absence of the cleaving by the restriction enzyme depends on whether the mutation of the particular base exists or not; and then, the resultant is electrophoresed to detect whether the products obtained by the amplification have been cleaved or not (see, for example, Genet. Mol. Res., 2010, Vol. 9, No. 1, pp. 34-40).
In addition, a method, in which a region containing a mutation is amplified by a PCR method; thereafter, melting curve analysis is carried out using a nucleic acid probe that has been labeled with a fluorescent dye; and, based on the results of the melting curve analysis, the mutation in the base sequence is analyzed, is also known (see, for example, Japanese Patent Application Laid-Open (JP-A) No. 2002-119291).
However, in the PCR-RFLP method, it is necessary to carry out a PCR reaction, and thereafter collect the amplification products, and treat them with a restriction enzyme. Therefore, there may be a risk that the amplification products may contaminate the following reaction system, and this may cause a false-positive or false-negative result. In addition, since the restriction enzyme treatment is carried out after the completion of PCR and then the resultant is electrophoresed, it may take a very long time until the detection. Furthermore, this method is hard to automate due to complex operations thereof.
Therefore, it has been demanded to further develop a technique for detecting polymorphism in the ABCG2 gene.